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United States Patent |
5,233,029
|
Hammann
,   et al.
|
August 3, 1993
|
Elaiophylin derivatives and a process for the preparation thereof
Abstract
The invention relates to elaiophylin derivatives of the formula I, II and
III
##STR1##
in which the substituents R(1), R(2), R(3) and R(3)' have the specified
meanings. The compounds have antibacterial and antiviral activity.
Inventors:
|
Hammann; Peter (Taunus, DE);
Kretzschmar; Gerhard (Eschborn, DE)
|
Assignee:
|
Hoechst Aktiengesellschaft (Frankfurt am Main, DE)
|
Appl. No.:
|
607285 |
Filed:
|
October 31, 1990 |
Foreign Application Priority Data
Current U.S. Class: |
536/7.1; 536/16.8; 548/251; 548/255; 548/263.2; 548/266.2; 548/305.4; 548/306.1; 548/311.1; 548/313.1; 548/517; 549/267 |
Intern'l Class: |
C07H 017/08; C07D 409/02; C07D 257/02; C07D 321/00 |
Field of Search: |
549/267,60
548/251,255,263.2,266.6,336,397,517
514/450
536/7.1,16.8
|
References Cited
U.S. Patent Documents
5011827 | Apr., 1991 | Kretzschmar et al. | 536/7.
|
Foreign Patent Documents |
0361467 | Apr., 1990 | EP.
| |
Primary Examiner: Ivy; C. Warren
Assistant Examiner: Owens; A. A.
Attorney, Agent or Firm: Finnegan, Henderson, Farabow, Garrett & Dunner
Parent Case Text
This is a division of application Ser. No. 07/407,617, filed Sep. 15, 1989
now U.S. Pat. No. 4,985,451.
Claims
We claim:
1. A compound III
##STR17##
wherein R(1) and R(2) denote, identically or differently, hydrogen, a
radical of the formula V or V'
##STR18##
wherein n equals 1-3 and R(5) is equal to (C.sub.1 -C.sub.5)-alkyl,
(C.sub.2 -C.sub.15)-alkenyl, (C.sub.2 -C.sub.15)-alkynyl, (C.sub.3
-C.sub.9)-cycloalkyl, phenyl, naphthyl, furyl, thienyl, unsubstituted or
substituted by halogen, NO.sub.2, CH, OH, COOH, (C.sub.1 -C.sub.4)-alkyl
or (C.sub.1 -C.sub.4)-alkoxy,
and in the case where R(1) equals H, and R(2) also equals H, a radical of
the formula VI
SO.sub.2 R(6) VI
wherein R(6) is equal to (C.sub.1 -C.sub.10)-alkyl, phenyl, or tolyl.
2. A process for the preparation of a compound I of the following formula:
##STR19##
where the C--C double bonds in the macrodiolide ring of said compound I
can also be hydrogenated, wherein, in said formula:
a) R(3) and R(3)', being identical or different, are a radical of the
formula IV
--SR(4)
where R(4) is hydrogen, (C.sub.1 -C.sub.10)-alkyl which is unsubstituted
or substituted by OH or COOH, (C.sub.2 -C.sub.10)-alkenyl, (C.sub.3
-C.sub.8)-cycloalkyl, pyrrolyl, benzopyrrolyl, imidazolyl, benzimidazolyl,
triazolyl, tetrazolyl, phenyl, with the aromatic or heteroaryl radicals
being unsubstituted or substituted once or twice by (C.sub.1
-C.sub.4)-alkyl, (C.sub.1 -C.sub.4)-alkoxy, (C.sub.1
-C.sub.4)-alkylcarbonyl, carboxyl, F, Cl, Br, I, NO.sub.2, or CN;
b) R(3) is L-deoxyfucose and R(3)' is as defined under a); or
c) if the C--C double bonds in the macrodiolide ring are hydrogenated:
R(3) and R(3)' are hydrogen or
R(3) is L-deoxyfucose and R(3)' is hydrogen;
said process comprising the steps wherein:
a) elaiophylin
##STR20##
is reacted in a mixture of water, alcohol, and an inert organic solvent
with a base having a pH of 8 to 11 to give a compound II
##STR21##
or III
##STR22##
and the compound II or III, which is obtained in this way and in which
R(1)=R(2)=H, is reacted with a thiol ether of the formula IV
HSR(4)
in which R(4) is as defined above, to give a compound I, or in the case in
which compound II or III is obtained in this way and in which R(1)=R(2)=H
is completely hydrogenated with hydrogen, a compound I wherein
R(3)=L-deoxyfucose and R(3)'=hydrogen is obtained from compound II, and a
compound I wherein R(3)=R(3)'=hydrogen is obtained from compound III; or
said process comprises the steps wherein:
b) a compound II
##STR23##
or III
##STR24##
in which R(1)=R(2)=H is reacted with a thiol of the formula
HSR(4)
in which R(4) is as defined above, in which case a compound I wherein
R(3)=L-deoxyfucose and R(3)'=SR(4) is obtained from compound II; and a
compound I wherein R(3)=R(3)'=SR(4) is obtained from compound III; or said
process comprises the steps wherein:
c) a compound II or III in which the C--C double bonds in the macrodiolide
ring can also be hydrogenated and in which R(1)=R(2)=H is completely
hydrogenated, in which case a compound I with R(3)=L-deoxyfucose and
R(3)'=hydrogen is obtained from compound II, and a compound I with
R(3)=R(3)'=hydrogen is obtained from compound III.
3. A process for the preparation of a compound I according to claim 2
wherein said alcohol is selected from the group consisting of methanol,
ethanol and isopropanol.
4. A process for the preparation of a compound I according to claim 2
wherein said inert organic solvent is selected from the group consisting
of chloroform, ethyl acetate, tetrahydrofuran, methylene chloride and
dioxane.
5. A process for the preparation of a compound I according to claim 2
wherein said base is selected from the group consisting of alkali metal
and alkaline earth metal hydroxides, carbonates and bicarbonates.
6. A process for the preparation of a compound I according to claim 2
wherein the elaiophylin is reacted in a mixture of water, ethanol, ethyl
acetate and sodium bicarbonate or potassium bicarbonate, said mixture
having a pH of 8 to 11.
Description
The invention relates to elaiophylin derivatives and to the preparation
thereof and the use thereof as pharmaceuticals, in particular as
pharmaceuticals having antibacterial and antiviral activity. Elimination
of the 2,6-deoxyfucose from elaiophylin gives rise to difficulties because
of the instability of the molecule both under acid and under basic
conditions, in this connection see D. Seebach, Liebigs Ann. Chem. 1986,
page 1281 and W. Keller-Schierlein, Helv. Chim. Acta 64, 407 (1981), S.
Takahashi Chem. Pharm. Bull. 15, 1651, 1657, 1726 (1976). The invention
now makes available for the first time conditions under which it is
possible to eliminate the deoxyfucose from elaiophylin
##STR2##
by controlled basic reaction conditions to give the unsymmetrical enones
II
##STR3##
and the symmetrical enones III
##STR4##
in high yields.
Addition of thio compounds to III results in elaiophylin derivatives I
##STR5##
and hydrogenation results in the corresponding octahydro compounds I with
R(3)=H and R(3)'=H or R(3)=L-deoxyfucose and R(3)'=H.
Hence the invention relates to compounds I
##STR6##
where the C--C double bonds in the macrodiolide ring of the compound of
the formula I can also be hydrogenated, the meanings in this formula I
being:
a) R(3)', identical or different, a radical of the formula IV
--SR(4) IV
with R(4) equal to hydrogen, (C.sub.1 -C.sub.10)-alkyl which is
unsubstituted or substituted by OH or COOH, (C.sub.2 -C.sub.10)-alkenyl,
(C.sub.3 -C.sub.8)-cycloalkyl, pyrrolyl, benzopyrrolyl, imidazolyl,
benzimidazolyl, triazolyl, tetrazolyl, phenyl,with the aromatic or
heteroaryl radicals being unsubstituted or substituted once or twice by
(C.sub.1 -C.sub.4)-alkyl, (C.sub.1 -C.sub.4)-alkoxy, (C.sub.1
-C.sub.4)-alkylcarbonyl, carboxyl, F, Cl, Br, I, NO.sub.2 or CN.
b) R(3) L-deoxyfucose and R(3)' as defined under a).
c) If the C--C double bonds in the macrodiolide ring are hydrogenated:
R(3), and R(3)' hydrogen or
R(3) L-deoxyfucose and R(3)' hydrogen.
Preferred compounds I are those in which R(3) and R(3)' represent a radical
of the formula IV with R(4) equal to hydrogen, (C.sub.1 -C.sub.5)-alkyl
which is unsubstituted or substituted by OH or COOH, (C.sub.2
-C.sub.5)-alkenyl, (C.sub.5 -C.sub.8)-cycloalkyl, pyrrolyl, benzopyrrolyl,
imidazolyl, phenyl, which is unsubstituted or substituted by (C.sub.1
-C.sub.4)-alkyl, (C.sub.1 -C.sub.4)-alkoxy, F, Cl, Br, NO.sub.2 or CN.
Particularly preferred compounds I are those in which R(3) is the
L-deoxyfucosyl group and R(3)' is a radical SR(4) IV as defined above.
Very particularly preferred compounds I are those in which the macrodiolide
ring is hydrogenated and in which R(3) is the L-deoxyfucosyl radical or
hydrogen and R(3)' is hydrogen.
The invention also relates to a process for the preparation of a compound
I, according to which
a) elaiophylin
##STR7##
is reacted with a base in aqueous solvent at a pH of 8 to 11 to give a
compound II
##STR8##
or III
##STR9##
and the compound II or III which is obtained in this way and in which
R(1)=R(2)=H is reacted with a thiol either of the formula IV
HSR(4) IV
in which R(4) is as defined in claim 1, to give a compound I, or the
compound II or III which is obtained in this way and which has R(1)=R(2)=H
is completely hydrogenated, in which case compounds I with
R(3)=L-deoxyfucose and R(3) equals hydrogen are obtained from compounds
II, and compounds I with R(3)=R(3)'=hydrogen are obtained from compounds
III with R(1)=R(2)=hydrogen; or
b) a compound II
##STR10##
or III
##STR11##
in which R(1)=R(2)=H is reacted with a thiol of the formula
HSR(4)
in which R(4) is as defined in claim 1, in which case the compound I with
R(3)=L-deoxyfucose and R(3)'=SR(4) IV is obtained from compound II; and
the compound I with R(3)=R(3)'=SR(4) from compound III; or
c) a compound II or III in which the C--C double bonds in the macrodiolide
ring can also be hydrogenated and which has R(1)=R(2)=H is completely
hydrogenated, in which case compounds I with R(3)=L-deoxyfucose and R(3)'
equals hydrogen are obtained from compounds II, and compounds I with
R(3)=R(3)'=hydrogen are obtained from compounds III with
R(1)=R(2)=hydrogen.
The invention additionally relates to compounds III
##STR12##
in which the following substituents have the following meaning
R(1) and R(2), identical or different, hydrogen, a radical of the formula V
or V'
##STR13##
with n=1-3 and R(5) equal to (C.sub.1 -C.sub.5)-alkyl, (C.sub.2
-C.sub.15)-alkenyl, (C.sub.2 -C.sub.15)-alkynyl, (C.sub.3
-C.sub.9)-cycloalkyl, phenyl, naphthyl, furyl, thienyl, unsubstituted or
substituted by F, Cl, Br, I, NO.sub.2, CN, OH, COOH, (C.sub.1
-C.sub.4)-alkyl or (C.sub.1 -C.sub.4)-alkoxy, and in the case where R(1)
equals H, R(2) also equals H, a radical of the formula VI
SO.sub.2 R(6) VI
with R(6) equal to (C.sub.1 -C.sub.10)-alkyl, phenyl, tolyl, and in the
case where R(1) equals H, R(2) also equals H.
The invention is also directed at compounds II
##STR14##
in which the following substituents have the following meaning
R(1) and R(2), identical or different, hydrogen, a radical of the formula V
or V'
##STR15##
with n=1-3 and R(5) equal to (C.sub.1 -C.sub.5)-alkyl, (C.sub.2
-C.sub.15)-alkenyl, (C.sub.2 -C.sub.15)-alkynyl, (C.sub.3
-C.sub.9)-cycloalkyl, aryl, heteroaryl, unsubstituted or substituted by
halogen, NO.sub.2, CN, OH, COOH, (C.sub.1 -C.sub.4)-alkyl or (C.sub.1
-C.sub.4)-alkoxy, and in the case where R(1) equals H, R(2) also equals H,
a radical of the formula VI
SO.sub.2 R(6) VI
with R(6) equal to (C.sub.1 -C.sub.10)-alkyl, phenyl, tolyl, and in the
case where R(1) equals H, R(2) also equals H.
Also preferred are intermediate compounds II and III in which R(1) and R(2)
are hydrogen or, independently of one another, represent a radical of the
formula V in which R(5) has the meaning (C.sub.1 -C.sub.5)-alkyl, (C.sub.5
-C.sub.6)-cycloalkyl or phenyl, and in the case where R(1) is hydrogen,
R(2) is also hydrogen. Likewise preferred are compounds II and III in
which R(1) represents a radical of the formula VI with R(6)=(C.sub.1
-C.sub.5)-alkyl, phenyl or tolyl. All these compounds possess as
antibacterial activity against bacteria pathogenic for humans, as well as
antiviral activity.
The compounds I, II and III are obtained by various processes.
a. Elaiophylin is reacted in a solvent mixture with a base which has a pH
of 8 to 11, in which case the L-deoxyfucose is eliminated by
.beta.-elimination to give compound II (R(1), R(2)=H) and compound III
(R(1), R(2)=H).
b. If compound II or III with R(1)=R(2)=hydrogen is reacted with a thiol of
the formula HSR(4) as defined above, the corresponding compounds I are
obtained by nucleophilic addition onto the enone system. Thus, II gives
the compound I with R(3)=L-deoxyfucose and R(3)'=IV, and III gives the
compound I with R(3)=R(3)'=IV.
c. If compound II or III with R(1)=R(2)=H is completely hydrogenated with
hydrogen, the compound I with R(3)=L-deoxyfucose and R(3)'=hydrogen is
obtained from II, and the compound I with R(3)=R(3)'=hydrogen is obtained
from compound III (octahydrodideoxyelaiophylidene).
d. If compound II or III with R(1)=R(2)=hydrogen or R(1)=V or V' as defined
above, or R(1)=VI as defined above, and R(2)=hydrogen is reacted with a
compound of the formula VII or VII'
##STR16##
in which n and R(5) have the meaning specified under V and V', and Q
denotes chloride, bromide, imidazolide or acid anhydride, a compound with
R(1)=V or V' and R(2)=hydrogen or compounds with R(1), R(2)=V or V' are
obtained from compounds II or III with R(1)=R(2)=H, and compounds with
R(1)=V or V' and R(2)=V or V', in which R(1) and R(2) are identical or
different, are obtained from compounds II or III with R(1)=V and
R(2)=hydrogen.
e. If a compound II or III with R(1)=R(2)=hydrogen or R(1)=V or V' as
defined above, or R(1)=VI as defined above, and R(2)=hydrogen is reacted
with a compound of the formula ClSO.sub.2 R(6), compounds with R(1)=VI and
R(2)=hydrogen or R(1)=R(2)=VI are obtained from compound II or III with
R(1)=R(2)=hydrogen, or the derivatives with R(1)=V or V' and R(2)=VI are
obtained from compounds II or III with R(1)=V or V', R(2)=hydrogen, as
well as the derivatives with R(1)=VI and R(2), where R(1) and R(2) can be
identical or different, are obtained from R(1)=VI and R(2)=hydrogen.
Processes a to e are described in more detail hereinafter. It is possible
by variant a to eliminate the L-deoxyfucose side-chain from elaiophylin.
This results in compounds II and III with R(1)=R(2)=hydrogen. The yield of
II decreases and that of III increases with higher temperatures and longer
reaction times.
The best procedure is to suspend elaiophylin in a mixture of water, alcohol
and an inert organic solvent. Suitable alcohols are methanol, ethanol and
isopropanol, and suitable inert solvents are chloroform, ethyl acetate,
THF, methylene chloride.
The mixture of water, ethanol and ethyl acetate is preferred. The reaction
temperatures are between 20.degree. C. and the boiling point of the
reaction mixture. The reaction is preferably carried out under reflux. The
reaction times amount to 0.5 to 180 hours, preferably 1 to 24 hours. The
completion of the reaction is determined by thin-layer chromatography.
Bases which are used are alkali metal and alkaline earth metal hydroxides
and carbonates and bicarbonates. The pH of the reaction solution should be
between 7 and 14, preferably between 8 and 11. Sodium bicarbonate and
potassium bicarbonate at a pH of 8 to 11 are very particularly preferred.
The pH can be controlled with HCl or H.sub.2 SO.sub.4 or via an
appropriate buffer.
The elaiophylin required as starting substance can be prepared by known
preparation processes. That of German Patent Application P 3721722.4 is a
suitable example. This results in elaiophylin as a product of fermentation
of cultures of the strains DSM 4137 and DSM 3816.
In process b, compound II or III with R(1)=R(2)=hydrogen is dissolved in a
solvent such as alcohol or CHCl.sub.3, CH.sub.2 Cl.sub.2, THF, dioxane,
and reacted with equimolar amounts or with an up to 50-fold excess of
thiol of the formula HSR.sub.4 in the presence of a base. Especially
suitable bases are organic amines, preferably triethylamine, and alcohols
which are used are especially methanol, ethanol or isopropanol. If salts
M.sup.+- SR(4) are used directly, with M.sup.+ equal to sodium.sup.+ or
potassium.sup.+ for example, a suitable solvent is THF or dioxane. These
salts are then prepared in THF or dioxane from HSR(4) by addition of the
appropriate metal hydrides. The reaction temperatures are between
0.degree. C. and the boiling point of the solvent. Temperatures between
50.degree. C. and the boiling point are preferred. The reaction times are
0.1-180 hours, preferably 0.1 to 120 hours, depending on R(4).
The C--C double bonds of the macrodiolide ring as well as the enone double
bond in II and III (R.sub.1 =R.sub.2 =H) can be hydrogenated by process
variant c).
The best procedure for process variant c) is to react the elaiophylin
derivative II R.sub.1 /R.sub.2 =H or III R.sub.1 /R.sub.2 =H which is to
be hydrogenated and which is preferably dissolved in a solvent such as
methanol, ethanol, isopropanol or ethyl acetate, or a mixture of these
solvents or an aqueous mixture of these solvents, with hydrogen in the
presence of a conventional hydrogenation catalyst by hydrogenation
processes known from the literature. Examples of conventional
hydrogenation catalysts are elements of group 8 such as platinum,
palladium or else nickel, which are usually supported, for the purpose of
increasing the reactive surface, on active carbon, silica or alumina
supports, for example. If the reaction is carried out in an absolute
primary alcohol as solvent, apart from a hydrogenation of the C.dbd.C
double bonds there is also ketalization to give the C.sub.11 /C.sub.11
'-di-O-alkylene.
Depending on the catalyst used, the reaction can be carried out both
without and with an excess pressure of hydrogen, for example up to 1
atmosphere. The reaction temperatures are between 0.degree. C. and
40.degree. C., preferably at room temperature. The reaction times depend
on the batch size and the concentration of the compound to be reduced.
Hydrogenation processes of this type are described, for example, in
Organikum, Organisch Chemisches Grundpraktikum (Basic Techniques of
Organic Chemistry), 15th edition, VEB Deutscher Verlag der Wissenschaften,
Berlin, 1976, pages 359-371.
The hydroxyl groups in formula II and III can be esterified by process
variant d). Since the reaction rate for esterification at the 3', 4' and
15 position--that is to say for the R.sup.1 derivatives--is greater than
the esterification at the 9 position, the possibility of providing for
R.sup.1 and R.sup.2 both identical and different substituents opens up.
With higher temperatures and/or sufficiently long reaction times the OH
group in the 9 position is also esterified. However, this also means that
esterification in the 9 position is possible only after previous
esterification of the OH groups in the 3', 4' and 15 position. Thus, for
example, it is possible in this way for first the OH groups in the 3', 4'
and 15 position to be esterified and, subsequently, where appropriate
after isolation and purification of the product, for the OH group in the 9
position to be esterified in a second reaction corresponding to process
variant a.
The best procedure for process variant b) is to react a compound of the
formula II or III in which R.sup.1, R.sup.2 have the meanings specified
above, in equimolar amounts or in an up to 50-fold excess, where
appropriate in an inert aprotic solvent such as chloroform, methylene
chloride, tetrahydrofuran (THF), ethyl acetate or dioxane, with a compound
of the formula VII or VII' until the reaction is complete, where
appropriate in the presence of a base, preferably pyridine.
The temperatures for this reaction are between -70.degree. C. and
+100.degree. C., preferably when a solvent is used between the
solidification point and the boiling point of the solvent, in particular
between -70.degree. C. and +40.degree. C. The reaction times amount to 1
to 180 hours, preferably 1 to 48 hours, particularly preferably 1 to 8
hours. The completion of the reaction can be determined, for example, by
thin-layer chromatography (TLC monitoring).
The starting compounds for process variant d), which are compounds of the
formula VII and/or VII', can, where they cannot be bought, be prepared in
a straightforward manner by processes known from the literature. For
example, the acid chlorides are obtained by reacting the corresponding
carboxylic acid with thionyl chloride, PCl.sub.3 or PCl.sub.5. Processes
of this type are described, for example, in Gattermann/Wieland, "Die
Praxis des Organischen Chemikers" (Practical Organic Chemistry), 43rd
edition, Walter de Gruyter, Berlin, N.Y. 1982, pages 303 et seq.
The best procedure for process variant e) is to react an elaiophylin
derivative II or III in equimolar amounts or in an up to 50-fold excess
with sulfonyl halides of the formula Y--SO.sub.2 R.sup.6.
This reaction can also be carried out with addition of base where
appropriate. Examples of suitable bases are triethylamine, pyridine or
lutidine. One variant of process e) comprises using a suitable, preferably
inert solvent such as chloroform, methylene chloride, THF, ethyl acetate
or dioxane. It is also possible in this case for the excess of the
compounds listed above to be up to 50 times the amount.
The temperature for this reaction are between -70.degree. C. and
+100.degree. C., preferably when a solvent is used between the
solidification point and the boiling point of the solvent, in particular
between -70.degree. C. and -40.degree. C. The reaction times amount to 1
to 180 hours, preferably 1 to 48 hours, particularly preferably 1 to 8
hours. The completion of the reaction can be determined, for example, by
TLC monitoring.
The starting compounds for process variant e) can, where they cannot be
bought, be prepared in a straightforward manner by processes known from
the literature. For example, the sulfonyl halides of the formula
Y--SO.sub.2 R.sup.6 are obtained by radical reaction of alkanes with
chlorine and SO.sub.2 or by halogenation of aromatic compounds with
halogenosulfonic acid Y--SO.sub.3 H.
The antiviral activity of the compounds I, II and III according to the
invention was tested in cell cultures infected with test viruses. It
emerged from this that the derivatives according to the invention have an
exellent antiviral effect.
The compounds according to the invention are suitable, by reason of their
pharmacological properties, for the treatment of bacterial diseases and
viral diseases caused, for example, by HSV I, II (herpes simplex I or II
virus) or else picorna- and retroviruses such as HIV (human
immunodeficiency virus).
The invention therefore additionally relates to the use of the compounds of
the formula I, II and III according to the invention for the treatment and
prophylaxis of bacterial diseases or of herpes virus, picorna- and
retroviral diseases.
The compounds can be used as pharmaceuticals either alone or mixed with
physiologically tolerated auxiliaries or excipients. For this purpose they
can be administered orally in doses of 0.01-5.0 mg/kg/day, preferably
0.01-1.0 mg/kg/day or parenterally subcutaneously in doses of 0.001-2.5
mg/kg/day, preferably 0.001-1.0 mg/kg/day, in particular 0.005-0.2
mg/kg/day. Topical use is particularly preferred, in which case the
concentration of active substance in the ointments is 0.001-1%, preferably
0.01-0.1%. The dosage can also be increased in severe cases. However,
lower doses also suffice in many cases. The statements relate to an adult
weighing about 75 kg.
The invention additionally embraces the use of the compounds according to
the invention for the preparation of pharmaceuticals which are used for
the treatment and prophylaxis of the abovementioned diseases.
The invention further relates to pharmaceuticals which contain one or more
compounds of the formula I, II and/or III according to the invention.
The pharmaceuticals are prepared by processes which are known per se and
familiar to those skilled in the art. As pharmaceuticals, the
pharmacologically active compounds (=active substance) according to the
invention are used either as such or, preferably, in combination with
suitable pharmaceutical auxiliaries or excipients in the form of tablets,
coated tablets, capsules, suppositories, emulsions, suspensions or
solutions, with the content of active substance being up to about 95%,
preferably between 10 and 75%. On topical use, concentrations of active
substance of 0.001-1%, preferably 0.01-0.1%, suffice.
Examples of suitable auxiliaries and excipients for the desired
pharmaceutical formulation are, besides solvents, gel-formers, suppository
bases, tablet auxiliaries and other active substance vehicles, also
antioxidants, dispersing agents, emulsifiers, antifoam agents, flavorings,
preservatives, solubilizers or colorants.
The active substances can be administered orally, parenterally
(subcutaneously), topically or rectally, with topical application being
preferred. The active compounds are mixed with the additives suitable for
this purpose, such as excipients, stabilizers or inert diluents, and
converted by the customary methods into suitable dosage forms such as
tablets, coated tablets, hard gelatine capsules, aqueous, alcoholic or
oily suspensions or aqueous or oily solutions, creams or ointments.
Examples of inert excipients which can be used are gum arabic, magnesia,
magnesium carbonate, potassium phosphate. This formulation can be carried
out both as dry and as wet granules. Examples of suitable oily excipients
or solvents are vegetable or animal oils such as sunflower oil or fish
liver oil.
For subcutaneous or intravenous administration, the active compounds are
converted, if desired with the substances suitable for this purpose, such
as solubilizers, emulsifiers or other auxiliaries, into solution,
suspension or emulsion. Examples of suitable solvents are physiologically
saline or alcohols, for example ethanol, propanol, glycerol, as well as
sugar solutions such as glucose or mannitol solutions, or else a mixture
of the various solvents mentioned. The invention is explained in more
detail hereinafter by means of examples.
EXAMPLES
Process a
12 mmol of elaiophylin are heated in 120 ml of water, 80 ml of ethanol and
70 ml of ethyl acetate with 25 g of KHCO.sub.3 at 65.degree. C. The
mixture is then concentrated in vacuo, and the remaining aqueous phase is
extracted with ethyl acetate (4.times.150 ml). Drying over sodium sulfate
is followed by concentration in vacuo. The remaining residue is
chromatographed on silica gel (elution with CHCl.sub.3 /CH.sub.3 OH 40:1).
Process b
1 mmol of elaiophylin derivative is stirred in 30 ml of ethanol with 300 ml
of ethanethiol and 300 ml of triethylamine at 60.degree. C. for 1 hour.
Addition of 50 ml of water is followed by extraction with ethyl acetate
(3.times.50 ml). The ethyl acetate phase is washed with 30 ml of water
(3.times.) and dried over sodium sulfate. Evaporation in vacuo is followed
by chromatography of the residue on silica gel (ethyl acetate/hexane 1:4).
Process c
1 mmol of elaiophylin derivative 1 or 5 is dissolved in 30 ml of ethyl
acetate, and 10% by weight of Pd/animal charcoal is added. Hydrogenation
is carried out at room temperature while stirring in a closed
hydrogenation apparatus maintaining a slight excess pressure of up to 0.2
bar of hydrogen. After 3 hours, the mixture is filtered and the solvent is
removed. Chromatography on silica gel with ethyl acetate/hexane 1:3 to 3:1
yields the corresponding products.
Process d
1 mmol of elaiophylin derivative is dissolved in 20 ml of pyridine and
stirred with 10 ml of acid anhydride at room temperature for 24 hours.
Hydrolysis with 60 ml of water is followed by extraction with ethyl
acetate (3.times.30 ml). The organic phase is washed with 10 ml of 1N HCl
and 20 ml of water and dried over sodium sulfate. Evaporation to dryness
in vacuo in a rotary evaporator is followed by chromatography on silica
gel (elution CHCl.sub.3 /CH.sub.3 OH from 30:1 to 10:1).
Process e
1 mmol of elaiophylin derivative is dissolved in 20 ml of pyridine and
stirred with 10 mmol of methanesulfonyl chloride at room temperature for
one hour. Hydrolysis with 60 ml of water is followed by working up as in
process d.
______________________________________
Physicochemical data on the compounds prepared
melting
No. MNa.sup.+
point .sup.13 C in CDCl.sub.3
______________________________________
1 898 876,3 202,6, 168,4, 168,0, 148,2, 145,4,
145,3, 145,1, 144,9, 131,6, 131,4,
122,2, 122,1, 100,29, 94,2
2 1067 1045,6
3 1359 1337,6
4 1901 1379,5
5 751 728.9 202,6, 168,0, 148,3, 145,2, 245,0,
131,6, 131,4, 122,3, 76,8,
72,2, 68,9
6 919 897,1 200,4, 170,3, 170,1, 167,4, 146,0,
145,8, 144,4, 132,4, 131,7, 122,0,
75,2, 74,4,
7 959 937,2 202,23, 168,9, 167,9, 165,9, 150,0,
149, 149,3, 76,8, 72,8, 72,0
8 907 885,1 202,0, 168,0, 145,4, 145,1, 143,8,
132,8, 131,4, 122,2 80,9, 76,9,
71,9
9 891* 852,2 168,5, 195,4, 195,0, 131,5, 122,0,
99,2, 76,9, 70,1, 68,3
10 907* 884,2 168,5, 145,3, 145,0, 131,5, 122,0,
99,1, 76,9, 70,8, 68,3, 62,4,
11 763 741,06 Schmp. 152-153.degree. C.
12 908 886.2
______________________________________
*MK.sup.
__________________________________________________________________________
Examples Reaction
No. Formula
R.sub.1
R.sub.2
R.sub.3,
R.sub.3
Synth. from Reagent
time in h
Process
Yield
__________________________________________________________________________
1 II H H -- -- ELA
KHCO.sub.3
6 a 44%
2 II Ac Ac
-- -- 1 Ac.sub.2 O
24 d 85%
3 II Bz H -- -- 1 Bz.sub.2 O
24 d 72%
4 II Bz Ac
-- -- 3 Ac.sub.2 O
24 d 70%
5 III H H -- -- ELA
KHCO.sub.3
14 a 75%
6 III Ac Ac
-- -- 5 Ac.sub.2 O
24 d 85%
7 III Bz H -- -- 5 Bz.sub.2 O
24 d 78%
8 III CH.sub.3 SO.sub.2
H -- -- 5 CH.sub.3 SO.sub.2 Cl
11 e 92%
9 I -- --
SCH.sub.2 CH.sub.3
SCH.sub.2 CH.sub.3
5 CH.sub.3 CH.sub.2 SH
1 b 88%
10 I -- --
SCH.sub.2 CH.sub.2 OH
SCH.sub.2 CH.sub.2 OH
5 HOCH.sub.2 CH.sub.2 SH
1 b 86%
11 I -- --
H H 5 Pd/H.sub.2
3 c 92%
hydro-
genated
12 I -- --
H L-Deoxy-
5 Pd/12 3 c 94%
hydro- fucose
genated
__________________________________________________________________________
Ac = Acetyl
Bz = Benzoyl
Antiviral activity
Antiviral activity in cell cultures
The test substances were dissolved in cell culture medium (Dulbecco's MEM)
and introduced in a series of geometrical dilutions with a factor of 3
into 100 .mu.l of cell culture medium in standard microtiter plates.
Subsequently added were 100 .mu.l of a suspension of HeLa or Vero cells in
medium containing 5% fetal calf serum in a cell density of
2.times.10.sup.5 cells/ml. The mixtures were infected with 50 .mu.l of the
particular test virus in a suspension which was adjusted such that the
cells showed a cytopathogenic effect within 72 h. Evaluation was carried
out by microscopic inspection of the cell lawn and photometric measurement
of the uptake of neutral red (Finther staining test). The MIC was taken to
be the concentration of the product (.mu.g/ml) at which about 50% of the
cells survived the infection (MIC=minimum inhibitory concentration).
Table 1 shows the effect (MIC data in .mu.g/ml) of various compounds
according to the invention against the following viruses:
Herpes simplex virus I, herpes simplex virus II
TABLE 1
______________________________________
Compound Herpes Herpes
No. I II
______________________________________
1 4.94 4.94
9 44.4 14.8
10 4.94 1.65
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